Identification, expression and antigenic analysis of recombinant hemagglutinin proteins of canine distemper virus.
Canine distemper (CD) is a widely distributed disease of dogs, caused by the canine distemper virus (CDV). In the present study, the gene encoding the hemagglutinin (H) protein of a CDV isolate from central Taiwan was sequenced and compared with other strains. Sequence variations were noticed in the H gene from the field CDV strain that had previously been implicated in the increasing incidence of CD. To establish a serology-based diagnostic test, the full-length H protein, as well as five deletion mutants of a recombinant H protein of the local isolate, were produced using an E. coli expression system. Three truncated recombinant proteins with relatively high expression levels, designated HM3, HM4 and HM5, were used as antigens to examine their reactivity with canine sera. By using three negative sera and 17 CD-positive sera, the high specificity of recombinant H proteins was observed by ELISA. In addition, immunoblotting demonstrated that all three purified recombinant proteins exhibit an antigenic property recognized by the serum of a CD-suspected dog.
Chan
Kun-Wei
KW
Department of Veterinary Medicine College of Veterinary Medicine, National Chung-Hsing University, Taichung, Taiwan.
Hsieh
Hsien-Hua
HH
Wang
Hsien-Chi
HC
Lee
Ya-Jane
YJ
Sung
Ming-Hua
MH
Wong
Min-Liang
ML
Hsu
Wei-Li
WL
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D004216:Distemper / Q000175:diagnosis* / Q000821:virology; D004217:Distemper Virus, Canine / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay; D017353:Gene Deletion; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015151:Immunoblotting; D008969:Molecular Sequence Data; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D017422:Sequence Analysis, DNA
D000914:Antibodies, Viral; D006389:Hemagglutinins, Viral; D011994:Recombinant Proteins